Microarray-based assay for the detection of genetic variations of structural genes of West Nile virus.
Adaptation through fixation of spontaneous mutations in the viral genome is considered to be one of the important factors that enable recurrent West Nile virus (WNV) outbreaks in the U.S. Genetic variations can alter viral phenotype and virulence, and degrade the performance of diagnostic and screening assays, vaccines, and potential therapeutic agents. A microarray assay was developed and optimized for the simultaneous detection of any nucleotide mutations in the entire structural region of WNV in order to facilitate public health surveillance of genetic variation of WNV. The DNA microarray consists of 263 oligonucleotide probes overlapping at half of their lengths which have been immobilized on an amine-binding glass slide. The assay was validated using 23 WNV isolates from the 2002-2005 U.S. epidemics. Oligonucleotide-based WNV arrays detected unambiguously all mutations in the structural region of each one of the isolates identified previously by sequencing analysis, serving as a rapid and effective approach for the identification of mutations in the WNV genome.
Grinev
Andriyan
A
Laboratory of Molecular Virology, Division of Emerging and Transfusion Transmitted Diseases, Rockville, MD 20852, USA. Andriyan.Grinev@fda.hhs.gov
Daniel
Sylvester
S
Laassri
Majid
M
Chumakov
Konstantin
K
Chizhikov
Vladimir
V
Rios
Maria
M
D000818:Animals; D002522:Chlorocebus aethiops; D014644:Genetic Variation; D046228:Microarray Analysis / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes / Q000235:genetics; D017354:Point Mutation; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000235:genetics*; D014902:West Nile virus / Q000235:genetics*
D015345:Oligonucleotide Probes; D015678:Viral Structural Proteins